Cloning,expression, and characterization of catechol 1,2-dioxygenase from a phenol-degrading Candida tropicalis JH8 strain |
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Authors: | Yan Long Sheng Yang Li Cheng |
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Institution: | 1. College of Life Sciences, Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), State Key Laboratory of Virology, Wuhan University, Wuhan, China;2. Hubei Provincial Cooperative Innovation Center of Industrial Fermentation, Wuhan University, Wuhan, China;3. College of Life Sciences, Hubei University, Wuhan, China |
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Abstract: | The sequence cato encoding catechol 1,2-dioxygenase from Candida tropicalis JH8 was cloned, sequenced, and expressed in Escherichia coli. The sequence cato contained an ORF of 858?bp encoding a polypeptide of 285?amino acid residues. The recombinant catechol 1,2-dioxygenase exists as a homodimer structure with a subunit molecular mass of 32 KD. Recombinant catechol 1,2-dioxygenase was unstable below pH 5.0 and stable from pH 7.0 to 9.0; its optimum pH was at 7.5. The optimum temperature for the enzyme was 30°C, and it possessed a thermophilic activity within a broad temperature range. Under the optimal conditions with catechol as substrate, the Km and Vmax of recombinant catechol 1,2-dioxygenase were 9.2?µM and 0.987?µM/min, respectively. This is the first article presenting cloning and expressing in E. coli of catechol 1,2-dioxygenase from C. tropicalis and characterization of the recombinant catechol 1,2-dioxygenase. |
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Keywords: | Candida tropicalis JH8 catechol 1 2-dioxygenase cloning and expression enzymatic properties |
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